Oxidoreductases;
Acting on paired donors, with incorporation or reduction of molecular oxygen;
With NADH or NADPH as one donor, and incorporation of one atom of oxygen into the other donor
BRITE hierarchy

toluene,NADH:oxygen oxidoreductase (2,3-dihydroxylating)

(1) toluene + NADH + H+ + O2 = 2-methylphenol + NAD+ + H2O [RN:R03560];
(2) 2-methylphenol + NADH + H+ + O2 = 3-methylcatechol + NAD+ + H2O [RN:R03608]

R03560 R03608
Reaction

toluene [CPD:C01455];
NADH [CPD:C00004];
H+ [CPD:C00080];
O2 [CPD:C00007];
2-methylphenol

The enzyme, characterized from the bacterium Burkholderia cepacia, belongs to a class of nonheme, oxygen-dependent diiron enzymes. It contains a hydroxylase component with two binuclear iron centers, an NADH-oxidoreductase component containing FAD and a [2Fe-2S] iron-sulfur cluster, and a third component involved in electron transfer between the hydroxylase and the reductase. The enzyme dihydroxylates its substrate in two sequential hydroxylations, initially forming 2-methylphenol, which is hydroxylated to 3-methylcatechol.

EC 1.14.13.243 created 2019

1  [PMID:7578004]

Newman LM, Wackett LP

Purification and characterization of toluene 2-monooxygenase from Burkholderia cepacia G4.

Biochemistry 34:14066-76 (1995)
DOI:10.1021/bi00043a012

2  [PMID:9925593]

Yeager CM, Bottomley PJ, Arp DJ, Hyman MR

Inactivation of toluene 2-monooxygenase in Burkholderia cepacia G4 by alkynes.

Appl Environ Microbiol 65:632-9 (1999)
DOI:10.1128/AEM.65.2.632-639.1999

3  [PMID:11751810]

Canada KA, Iwashita S, Shim H, Wood TK

Directed evolution of toluene ortho-monooxygenase for enhanced 1-naphthol synthesis and chlorinated ethene degradation.

J Bacteriol 184:344-9 (2002)
DOI:10.1128/jb.184.2.344-349.2002